论文部分内容阅读
用与牛血清白蛋白偶联的南方水稻黑条矮缩病毒(Southern rice black-streaked dwarf virus,SRBSDV)衣壳蛋白的C端12个氨基酸多肽为抗原免疫BALB/c小鼠,经细胞融合、筛选、克隆,获得2株能稳定传代并分泌抗SRBSDV和水稻黑条矮缩病毒(Rice black-streaked dwarf virus,RBSDV)单克隆抗体(MAb)的杂交瘤细胞株3F1、5G1。3F1、5G1单克隆抗体腹水间接ELISA效价达10-6,抗体类型及亚类均为IgG1,kappa链。Western blot分析表明,2株单克隆抗体均与SRBSDV和RBSDV的外壳蛋白亚基有特异反应。利用单克隆抗体3F1建立的dot-ELISA检测方法能准确、特异、灵敏地检测田间稻飞虱及水稻样品中的SRBSDV和RBSDV。SRBSDV和RBSDV单克隆抗体的制备及检测方法的建立为水稻黑条矮缩病的诊断、预测预报及科学防控提供了技术支撑。
BALB / c mice were immunized with the 12 amino acid peptide of the C terminal of the southern rice black-streaked dwarf virus (BSBSDV) coupled with bovine serum albumin, After screening and cloning, two hybridoma cell lines, 3F1, 5G1.3F1 and 5G1, which can stably passage and secrete anti-SRBSDV and rice black-streaked dwarf virus monoclonal antibody (MAb) Indirect ELISA titer of clonal antibody ascites was 10-6, and the antibody types and subclasses were IgG1, kappa chain. Western blot analysis showed that both monoclonal antibodies reacted specifically with the coat protein subunits of SRBSDV and RBSDV. The dot-ELISA assay established by monoclonal antibody 3F1 can accurately, specifically and sensitively detect SRBSDV and RBSDV in rice planthoppers and rice samples. The establishment of the monoclonal antibodies against SRBSDV and RBSDV and the establishment of their detection methods provide technical support for the diagnosis, prediction and prevention of rice black-streaked dwarf diseases.